Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

Detalhes bibliográficos
Autor(a) principal: Mendes, Beatriz Madalena Januzzi
Data de Publicação: 2002
Outros Autores: Boscariol, Raquel Luciana, Filho, Francisco de Assis Alves Mourão, Almeida, Weliton Antonio Bastos de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Pesquisa Agropecuária Brasileira (Online)
Texto Completo: https://seer.sct.embrapa.br/index.php/pab/article/view/6425
Resumo: The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 mM of acetosyringone. Genetic transformation was confirmed by performing ß-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.
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spelling Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orangeTransformação genética de laranja 'Hamlin' via AgrobacteriumCitrus sinensis; epicotyls; seedlings; transgenics; breeding methodsCitrus sinensis; epicótilo; plântulas; transgênicos; métodos de melhoramento The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 mM of acetosyringone. Genetic transformation was confirmed by performing ß-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.O desenvolvimento e otimização de protocolos eficientes de transformação genética é essencial nos programas atuais de melhoramento de citros, tanto para porta-enxertos, como para copas de valor comercial. Plantas transgênicas de laranja 'Hamlin' (Citrus sinensis (L.) Osbeck) foram obtidas pela transformação genética de segmentos de epicótilo, coletados de plântulas germinadas in vitro, com Agrobacterium tumefaciens. Foram avaliados fatores que influenciam a eficiência da transformação genética, como: condições de incubação das plântulas utilizadas para coleta de explantes, tempo de inoculação com Agrobacterium e condições de co-cultivo. A regeneração de plantas a partir de segmentos de epicótilo ocorreu em alta freqüência, por organogênese direta. A maior porcentagem de plantas transgênicas foi obtida utilizando-se explantes coletados de plântulas germinadas no escuro e posteriormente transferidas, por 2-3 semanas, para condições de 16 horas de fotoperíodo, e infectados com Agrobacterium por um período de 15-45 minutos. As melhores condições de co-cultivo foram a incubação dos explantes no escuro, por três dias, em meio de cultura suplementado com 100 mM de acetoseringona. A transformação genética foi confirmada pelo teste histoquímico para ß-glucoronidase (GUS) e, posteriormente, pela amplificação de DNA, por PCR, para detecção dos genes nptII e GUS.Pesquisa Agropecuaria BrasileiraPesquisa Agropecuária BrasileiraMendes, Beatriz Madalena JanuzziBoscariol, Raquel LucianaFilho, Francisco de Assis Alves MourãoAlmeida, Weliton Antonio Bastos de2002-07-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.sct.embrapa.br/index.php/pab/article/view/6425Pesquisa Agropecuaria Brasileira; v.37, n.7, jul. 2002; 955-961Pesquisa Agropecuária Brasileira; v.37, n.7, jul. 2002; 955-9611678-39210100-104xreponame:Pesquisa Agropecuária Brasileira (Online)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPAenghttps://seer.sct.embrapa.br/index.php/pab/article/view/6425/3482info:eu-repo/semantics/openAccess2014-07-18T19:20:10Zoai:ojs.seer.sct.embrapa.br:article/6425Revistahttp://seer.sct.embrapa.br/index.php/pabPRIhttps://old.scielo.br/oai/scielo-oai.phppab@sct.embrapa.br || sct.pab@embrapa.br1678-39210100-204Xopendoar:2014-07-18T19:20:10Pesquisa Agropecuária Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false
dc.title.none.fl_str_mv Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
Transformação genética de laranja 'Hamlin' via Agrobacterium
title Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
spellingShingle Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
Mendes, Beatriz Madalena Januzzi
Citrus sinensis; epicotyls; seedlings; transgenics; breeding methods
Citrus sinensis; epicótilo; plântulas; transgênicos; métodos de melhoramento
title_short Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
title_full Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
title_fullStr Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
title_full_unstemmed Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
title_sort Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange
author Mendes, Beatriz Madalena Januzzi
author_facet Mendes, Beatriz Madalena Januzzi
Boscariol, Raquel Luciana
Filho, Francisco de Assis Alves Mourão
Almeida, Weliton Antonio Bastos de
author_role author
author2 Boscariol, Raquel Luciana
Filho, Francisco de Assis Alves Mourão
Almeida, Weliton Antonio Bastos de
author2_role author
author
author
dc.contributor.none.fl_str_mv

dc.contributor.author.fl_str_mv Mendes, Beatriz Madalena Januzzi
Boscariol, Raquel Luciana
Filho, Francisco de Assis Alves Mourão
Almeida, Weliton Antonio Bastos de
dc.subject.por.fl_str_mv Citrus sinensis; epicotyls; seedlings; transgenics; breeding methods
Citrus sinensis; epicótilo; plântulas; transgênicos; métodos de melhoramento
topic Citrus sinensis; epicotyls; seedlings; transgenics; breeding methods
Citrus sinensis; epicótilo; plântulas; transgênicos; métodos de melhoramento
description The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 mM of acetosyringone. Genetic transformation was confirmed by performing ß-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.
publishDate 2002
dc.date.none.fl_str_mv 2002-07-01
dc.type.none.fl_str_mv
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://seer.sct.embrapa.br/index.php/pab/article/view/6425
url https://seer.sct.embrapa.br/index.php/pab/article/view/6425
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://seer.sct.embrapa.br/index.php/pab/article/view/6425/3482
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Pesquisa Agropecuaria Brasileira
Pesquisa Agropecuária Brasileira
publisher.none.fl_str_mv Pesquisa Agropecuaria Brasileira
Pesquisa Agropecuária Brasileira
dc.source.none.fl_str_mv Pesquisa Agropecuaria Brasileira; v.37, n.7, jul. 2002; 955-961
Pesquisa Agropecuária Brasileira; v.37, n.7, jul. 2002; 955-961
1678-3921
0100-104x
reponame:Pesquisa Agropecuária Brasileira (Online)
instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
instacron:EMBRAPA
instname_str Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
instacron_str EMBRAPA
institution EMBRAPA
reponame_str Pesquisa Agropecuária Brasileira (Online)
collection Pesquisa Agropecuária Brasileira (Online)
repository.name.fl_str_mv Pesquisa Agropecuária Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
repository.mail.fl_str_mv pab@sct.embrapa.br || sct.pab@embrapa.br
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